Ojkic D, Nagy E
Department of Pathobiology, University of Guelph, Guelph, Ontario N1G 2W1, Canada.
Virology. 2001 May 10;283(2):197-206. doi: 10.1006/viro.2000.0890.
Two regions containing tandemly repeated sequences are present in the fowl adenovirus 9 (FAdV-9) genome. The longer repeat region (TR-2) is composed of 13 contiguous 135-bp-long direct repeats, the function of which is unknown. An infectious FAdV-9 genomic clone, constructed by homologous recombination in Escherichia coli, was used for engineering of recombinant viruses. The enhanced green fluorescence protein (EGFP) coding sequence was cloned in both rightward and leftward orientations so as to replace TR-2. Replication-competent recombinant FAdVs were recovered, demonstrating that TR-2 was dispensable for FAdV-9 propagation in vitro. The expression of EGFP in infected cells was demonstrated by fluorescence microscopy, immunoprecipitation, and RT-PCR.
禽腺病毒9型（FAdV-9）基因组中存在两个包含串联重复序列的区域。较长的重复区域（TR-2）由13个连续的135碱基对长的直接重复序列组成，其功能尚不清楚。通过在大肠杆菌中同源重组构建的具有感染性的FAdV-9基因组克隆被用于重组病毒的工程改造。增强型绿色荧光蛋白（EGFP）编码序列以正向和反向两种方向进行克隆，以取代TR-2。获得了具有复制能力的重组FAdV，表明TR-2对于FAdV-9在体外的增殖并非必需。通过荧光显微镜、免疫沉淀和RT-PCR证实了感染细胞中EGFP的表达。
